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Establishment and identification of an ovarian cancer cell strain stably expressing CDC50A
- Source :
- Jichu yixue yu linchuang, Vol 42, Iss 6, Pp 896-901 (2022)
- Publication Year :
- 2022
- Publisher :
- Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College., 2022.
-
Abstract
- Objective To establish a stable-expressing human cell cycle regulatory protein 50A (CDC50A) SKOV3 cell strain expressing vector containing CDC50A by molecular cloning. Methods CDC50A was introduced into the expression vector pLVX-IRES-GFP by molecular cloning method. The pLVX-CDC50A-GFP expression plasmid was packaged by lentivirus and stably transfected into SKOV3. SKOV3 cell strain with high-expressing CDC50A was established. The expression of Flag protein was analyzed by flow cytometry, Western blot and immunofluorescence. Results The construction of pLVX-CDC50A-GFP expressing vector was confirmed by gel electrophoresis and endonuclease digestion. The efficiency of infection was observed qualitatively under fluorescence microscope and detected quantitatively by flow cytometry as 86.5%. Flag-tag protein was detected by Western blot to present only in the positive group. Conclusions The SKOV3 cell strain was established by infecting with lentivirus of pLVX-CDC50A-GFP, laying a foundation for the mechanism study of CDC50A in ovarian cancer.
- Subjects :
- cdc50a
over-express
ovarian cancer
stable expression
Medicine
Subjects
Details
- Language :
- Chinese
- ISSN :
- 10016325
- Volume :
- 42
- Issue :
- 6
- Database :
- Directory of Open Access Journals
- Journal :
- Jichu yixue yu linchuang
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.be8a696b36a439687a9dc915339311a
- Document Type :
- article
- Full Text :
- https://doi.org/10.16352/j.issn.1001-6325.2022.06.003